EP3090053B1 - Künstliche nukleinsäuremoleküle - Google Patents
Künstliche nukleinsäuremoleküle Download PDFInfo
- Publication number
- EP3090053B1 EP3090053B1 EP14854873.8A EP14854873A EP3090053B1 EP 3090053 B1 EP3090053 B1 EP 3090053B1 EP 14854873 A EP14854873 A EP 14854873A EP 3090053 B1 EP3090053 B1 EP 3090053B1
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- EP
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- Prior art keywords
- ribosomal protein
- nucleic acid
- utr
- acid molecule
- protein
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Claims (25)
- Ein artifizielles Nukleinsäuremolekül, umfassenda. mindestens einen offenen Leserahmen (ORF);b. mindestens ein 3'-nicht-translatiertes Abschnittselement (3'-UTR-Element), umfassend eine Nukleinsäuresequenz, welche abgeleitet ist aus dem 3'-UTR eines ribosomalen Protein-Gens oder aus einer funktionalen Variante des 3'-UTR eines ribosomalen Protein-Gens, wobei die funktionale Variante mindestens 80% Identität mit dem 3'-UTR eines ribosomalen Protein-Gens aufweist; undc. ein 5'-UTR als ein zusätzliches 5'-terminales Element, wobei das 5'-UTR ein 5' TOP-UTR ist, welches das 5'-TOP-Motiv nicht umfasst.
- Artifizielles Nukleinsäuremolekül nach Anspruch 1, wobei das mindestens eine 3'-UTR-Element die Proteinproduktion von diesem artifiziellen Nukleinsäuremolekül verstärkt, stabilisiert und/oder verlängert.
- Artifizielles Nukleinsäuremolekül nach Anspruch 1 oder 2, wobei das 3'-UTR heterolog zum ORF ist.
- Das artifizielle Nukleinsäuremolekül nach einem der Ansprüche 1 bis 3, wobei das mindestens eine 3'-UTR-Element eine Nukleinsäure umfasst, welche abgeleitet ist aus der 3'-UTR eines eukaryotischen ribosomalen Protein-Gens, vorzugsweise aus dem 3'-UTR eines ribosomalen Protein-Gens eines Vertebraten, weiter bevorzugt aus dem 3'-UTR eines ribosomalen Protein-Gens eines Säugers, noch weiter bevorzugt aus dem 3'-UTR eines ribosomalen Protein-Gens eines Primaten, insbesondere eines humanen ribosomalen Protein-Gens oder eines ribosomalen Poteingens eines Nagers, insbesondere eines murinen ribosomalen Protein-Gens.
- Artifizielles Nukleinsäuremolekül nach Anspruch 1 bis 4, wobei
der offene Leserahmen (ORF) nicht für ein Reporter-Gen kodiert oder nicht von einem Reporter-Gen abgeleitet ist, wobei das Reporter-Gen vorzugsweise nicht ausgewählt ist aus der Gruppe, bestehend aus Globin-Proteinen (insbesondere Beta-Globin), Luciferase-Protein, Beta-Glucuronidase (GUS) und GFP-Protein oder Varianten davon, bevorzugt nicht EGFP, oder Varianten, die mindestens 70% Sequenzidentität mit einem Globin-Protein, einem Luciferase-Protein oder einem GFP-Protein aufweisen;
der offene Leserahmen (ORF) nicht für ein ribosomales Protein, vorzugsweise nicht für ein eukaryotisches ribosomales Protein kodiert, wobei das ribosomale Protein bevorzugt nicht ausgewählt ist aus dem ribosomalen Protein S6 (RPS6), dem ribosomalen Protein L36a-like (RPL36AL) oder dem ribosomalen Protein S16 (RPS16); oder
wobei das 3'-UTR nicht abgeleitet ist aus einem Virus. - Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 bis 5, wobei die 3'-UTR, vorzugweise das artifizielle Nukleinsäuremolekül, keine poly(A)-Sequenz oder kein Polyadenylierungssignal umfasst,
oder
wobei das 3'-UTR, vorzugsweise das artifizielle Nukleinsäuremolekül, weiterhin eine poly(A)-Sequenz und/oder ein Polyadenylierungssignal umfasst, wobei die poly(A)-Sequenz oder das Polyadenylierungssignal vorzugsweise 3'-terminal zum 3'-UTR-Element angeordnet ist. - Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 bis 6, wobei das mindestens eine 3'-UTR-Element eine Nukleinsäure-Sequenz umfasst, welche abgeleitet ist aus dem 3'-UTR einer Sequenz ausgewählt aus der Gruppe, bestehend aus ribosomalem Protein L9 (RPL9), ribosomalem Protein L3 (RPL3), ribosomalem Protein L4 (RPL4), ribosomalem Protein L5 (RPL5), ribosomalem Protein L6 (RPL6), ribosomalem Protein L7 (RPL7), ribosomalem Protein L7a (RPL7A), ribosomalem Protein L11 (RPL11), ribosomalem Protein L12 (RPL12), ribosomalem Protein L13 (RPL13), ribosomalem Protein L23 (RPL23), ribosomalem ProteinL18 (RPL18), ribosomalem Protein L18a (RPL18A), ribosomalem Protein L19 (RPL19), ribosomalem Protein L21 (RPL21), ribosomalem Protein L22 (RPL22), ribosomalem Protein L23a (RPL23A), ribosomalem Protein L17 (RPL17), ribosomalem Protein L24 (RPL24), ribosomalem ProteinL26 (RPL26), ribosomalem ProteinL27 (RPL27), ribosomalem Protein L30 (RPL30), ribosomalem Protein L27a (RPL27A), ribosomalem Protein L28 (RPL28), ribosomalem Protein L29 (RPL29), ribosomalem Protein L31 (RPL31), ribosomalem Protein L32 (RPL32), ribosomalem Protein L35a (RPL35A), ribosomalem Protein L37 (RPL37), ribosomalem Protein L37a (RPL37A), ribosomalem Protein L38 (RPL38), ribosomalem Protein L39 (RPL39), ribosomalem Protein, groß, P0 (RPLPO), ribosomalem Protein, groß, P1 (RPLP1), ribosomalem Protein, groß, P2 (RPLP2), ribosomalem Protein S3 (RPS3), ribosomalem Protein S3A (RPS3A), ribosomalem Protein S4, X-linked (RPS4X), ribosomalem Protein S4, Y-Iinked 1 (RPS4Y1), ribosomalem Protein S5 (RPS5), ribosomalem Protein S6 (RPS6), ribosomalem Protein S7 (RPS7), ribosomalem Protein S8 (RPS8), ribosomalem Protein S9 (RPS9), ribosomalem Protein S10 (RPS10), ribosomalem Protein S11 (RPS11), ribosomalem Protein S12 (RPS12), ribosomalem Protein S13 (RPS13), ribosomalem Protein S15 (RPS15), ribosomalem Protein S15a (RPS15A), ribosomalem Protein S16 (RPS16), ribosomalem Protein S19 (RPS19), ribosomalem Protein S20 (RPS20), ribosomalem Protein S21 (RPS21), ribosomalem Protein S23 (RPS23), ribosomalem Protein S25 (RPS25), ribosomalem Protein S26 (RPS26), ribosomalem Protein S27 (RPS27), ribosomalem Protein S27a (RPS27a), ribosomalem Protein S28 (RPS28), ribosomalem Protein S29 (RPS29), ribosomalem Protein L15 (RPL15), ribosomalem Protein S2 (RPS2), ribosomalem Protein L14 (RPL14), ribosomalem Protein S14 (RPS14), ribosomalem Protein L10 (RPL10), ribosomalem Protein L10a (RPL10A), ribosomalem Protein L35 (RPL35), ribosomalem Protein L13a (RPL13A), ribosomalem Protein L36 (RPL36), ribosomalem Protein L36a (RPL36A), ribosomalem Protein L41 (RPL41), ribosomalem Protein S18 (RPS18), ribosomalem Protein S24 (RPS24), ribosomalem Protein L8 (RPL8), ribosomalem Protein L34 (RPL34), ribosomalem Protein S17 (RPS17), ribosomalem Protein SA (RPSA), Ubiquitin A-52 residue ribosomalem Proteinfusionsproduct 1 (UBA52), Finkel-Biskis-Reilly murines Sarcoma-Virus (FBR-MuSV) Ubiquitously Expressed (FAU), ribosomalem Protein L22-like 1 (RPL22L1), ribosomalem Protein S17 (RPS17), ribosomalem Protein L39-like (RPL39L), ribosomalem Protein L10-like (RPL10L), ribosomalem Protein L36a-like (RPL36AL), ribosomalem Protein L3-like (RPL3L), ribosomalem Protein S27-like (RPS27L), ribosomalem Protein L26-like 1 (RPL26L1), ribosomalem Protein L7-like 1 (RPL7L1), ribosomalem Protein L13a Pseudogen (RPL13AP), ribosomalem Protein L37a Pseudogen 8 (RPL37AP8), ribosomalem Protein S10 Pseudogen 5 (RPS10P5), ribosomalem Protein S26 Pseudogen 11 (RPS26P11), ribosomalem Protein L39 Pseudogen 5 (RPL39P5), ribosomalem Protein, groß, PO Pseudogen 6 (RPLP0P6) und ribosomalem Protein L36 Pseudogen 14 (RPL36P14).
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 to 7, wobei das mindestens eine 3'-UTR-Element eine Nukleinsäure-Sequenz umfasst oder aus dieser besteht, welche eine Sequenzidentität von mindestens 80%, weiter bevorzugt von mindestens 90%, noch weiter bevorzugt von mindestens 95%, und noch weiter bevorzugt von mindestens 99% mit der Nukleinsäure-Sequenz, ausgewählt aus der Gruppe, bestehend aus SEQ ID Nr: 10 bis 205, aufweist.
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 to 8, weiterhin umfassend eine 5'-Cap-Struktur, eine poly(C)-Sequenz, einen Histon stem-loop und/oder ein IRES-Motiv.
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 to 9, wobei der Histon stem-loop eine Sequenz nach SEQ ID Nr: 5 umfasst.
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 to 10, wobei das artifizielle Nukleinsäuremolekül, vorzugsweise der offene Leserahmen, mindestens teilweise G/C-modifiziert ist, wobei vorzugsweise der G/C-Gehalt des offenen Leserahmens gegenüber dem offenen Leserahmen des Wildtyps erhöht ist, und/oder
wobei der offene Leserahmen eine kodon-optimierte Region umfasst, wobei vorzugsweise der offene Leserahmen kodon-optimiert ist. - Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1 bis 11, welches eine RNA ist, vorzugsweise ein mRNA-Molekül.
- Ein Vektor, umfassenda. einen offenen Leserahmen und/oder eine Klonierungsstelle;b. mindestens ein 3'-nicht-translatiertes Abschnittselement (3'-UTR-Element), umfassend eine Nukleinsäure-Sequenz, welche abgeleitet ist aus dem 3'-UTR eines ribosomalen Protein-Gens oder aus einer funktionalen Variante des 3'-UTR eines ribosomalen Protein-Gens, wobei diese funktionale Variante mindestens 80% Sequenzidentität mit dem 3'-UTR eines ribosomalen Protein-Gens aufweist; undc. ein 5'-UTR als ein zusätzliches 5'-terminales Element, wobei das 5'-UTR ein 5' TOP-UTR ist, welche das 5'-TOP-Motiv nicht umfasst.
- Vektor nach Anspruch 13, wobei der offene Leserahmen, das mindestens eine 3'-UTR-Element und/oder 5'-UTR charakterisiert ist/sind durch mindestens eines der Merkmale, wie in irgendeinem der Ansprüche 2 bis 12 in Hinblick auf das artifizielle Nukleinsäuremolekül definiert.
- Vektor nach Anspruch 13 oder 14, welcher ein Plasmid-Vektor oder ein viraler Vektor, vorzugsweise ein Plasmid-Vektor ist.
- Vektor nach einem der Ansprüche 13-15, welcher ein artifizielles Nukleinsäuremolekül nach einem der Ansprüche 2- 12 umfasst.
- Eine Zelle, umfassend das artifizielle Nukleinsäuremolekül nach einem der Ansprüche 1-12 oder einen Vektor nach einem der Ansprüche 13 bis 16.
- Eine pharmazeutische Zusammensetzung, umfassend das artifizielle Nukleinsäuremolekül nach einem der Ansprüche 1-12, den Vektor nach einem der Ansprüche 13 bis 16, oder die Zelle nach Anspruch 17.
- Pharmazeutische Zusammensetzung nach Anspruch 18, weiterhin umfassend ein oder mehr pharmazeutisch aktzeptable Vehikel, Streckmittel und/oder Exzipienten und/oder ein oder mehr Adjuvans/tien.
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1-12, Vektor nach einem der Ansprüche 13 bis 16, Zelle nach Anspruch 17, oder pharmazeutische Zusammensetzung nach Anspruch 18 oder 19 zur Verwendung als Medikament.
- Artifizielles Nukleinsäuremolekül nach einem der Ansprüche 1-12, Vektor nach einem der Ansprüche 13 bis 16, Zelle nach Anspruch 17, oder pharmazeutische Zusammensetzung nach Anspruch 18 oder 19 zur Verwendung als Vakzine oder zur Verwendung bei der Gentherapie.
- Ein in vitro-Verfahren zur Verstärkung, Stabilisierung und/oder Verlängerung der Proteinproduktion von einem artifiziellen Nukleinsäuremolekül, vorzugsweise von einem mRNA-Molekül oder einem Vektor, wobei das in vitro-Verfahren den Schritt der Assoziierung des Nukleinsäuremoleküls, vorzugsweise des mRNA-Moleküls oder des Vektors, mit einem 3'-UTR-Element umfasst, wobei das 3'-UTR-Element charakterisiert ist durch eines der Merkmale, wie in einem der Ansprüche 1 bis 12 in Hinblick auf das artifizielle Nukleinsäuremolelül definiert.
- Verwendung eines 3'-UTR-Element zur Verstärkung, Stabilisierung und/oder Verlängerung der Proteinproduktion von einem Nukleinsäuremolekül in vitro, vorzugsweise von einem mRNA-Molekül oder einem Vektor, wobei das 3'-UTR-Element charakterisiert ist durch mindestens eines der Merkmale, wie in einem der Ansprüche 1 bis 12 in Hinblick auf das artifizielle Nukleinsäuremolekül definiert.
- Ein Kit oder ein Kit von Teilen (kit of parts), umfassend ein artifizielles Nukleinsäuremolelül nach einem der Ansprüche 1-12, ein Vektor nach einem der Ansprüche 13 bis 16, eine Zelle nach Anspruch 17, und/oder eine pharmazeutische Zusammensetzung nach Anspruch 18 oder 19.
- Kit nach Anspruch 24, weiterhin umfassend Instruktionen zur Verwendung, Zellen zur Transfektion, ein Adjuvans, Mittel zur Verabreichung der pharmazeutischen Zusammensetzung, einen pharmazeutisch akzeptablen Träger und/oder eine pharmzeutisch akzeptable Lösung zur Auflösung oder Verdünnung des artifiziellen Nukleinsäuremoleküls, des Vektors, der Zellen oder der pharmazeutischen Zusammensetzung.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP18199987.1A EP3495486B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP14854873.8A EP3090053B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP20202943.5A EP3842537A1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| US16/030,018 US11254951B2 (en) | 2014-12-30 | 2018-07-09 | Artificial nucleic acid molecules |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP2013003946 | 2013-12-30 | ||
| PCT/EP2014/003480 WO2015101414A2 (en) | 2013-12-30 | 2014-12-30 | Artificial nucleic acid molecules |
| EP14854873.8A EP3090053B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
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| Application Number | Title | Priority Date | Filing Date |
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| EP18199987.1A Division EP3495486B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP18199987.1A Division-Into EP3495486B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP20202943.5A Division EP3842537A1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
Publications (2)
| Publication Number | Publication Date |
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| EP3090053A2 EP3090053A2 (de) | 2016-11-09 |
| EP3090053B1 true EP3090053B1 (de) | 2018-11-21 |
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| EP20202943.5A Pending EP3842537A1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP18199987.1A Active EP3495486B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP14854873.8A Active EP3090053B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
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| EP20202943.5A Pending EP3842537A1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
| EP18199987.1A Active EP3495486B1 (de) | 2013-12-30 | 2014-12-30 | Künstliche nukleinsäuremoleküle |
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| EP (3) | EP3842537A1 (de) |
Cited By (1)
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| CN114207134A (zh) * | 2019-03-25 | 2022-03-18 | 俄亥俄州国家创新基金会 | 工程化的mRNA序列及其用途 |
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| GB201810301D0 (en) * | 2018-06-22 | 2018-08-08 | Evox Therapeutics Ltd | Combinatorial gene therapy |
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- 2014-12-30 EP EP18199987.1A patent/EP3495486B1/de active Active
- 2014-12-30 EP EP14854873.8A patent/EP3090053B1/de active Active
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| CN114207134A (zh) * | 2019-03-25 | 2022-03-18 | 俄亥俄州国家创新基金会 | 工程化的mRNA序列及其用途 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP3495486A1 (de) | 2019-06-12 |
| EP3495486B1 (de) | 2020-12-16 |
| EP3090053A2 (de) | 2016-11-09 |
| EP3842537A1 (de) | 2021-06-30 |
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